ANTIOXIDANT AND ANTICANCER ACTIVITY OF

GARCINIA MORELLA FRUIT ON T CELL MURINE

LYMPHOMA

Bhaswati Choudhury

Senior Research Fellow

Drug Discovery Lab

Institute of Advanced Study in Science and

Technology

Guwahati , Assam, India

bhaswatichoudhury@ymail.com

T CELL LYMPHOMA

Lymphoma is the most common type of blood cancer. Two main forms

of lymphoma are Hodgkin’s lymphoma and Non Hodgkin’s lymphoma.

Lymphoma occurs when cells of the immune system called

lymphocytes, a type of white blood cell, grow and multiply

uncontrollably

Murine Dalton’s lymphoma (DLA)

is a Non Hodgkin’s transplantable

T cell lymphoma which can spread

beyond the lymphatic system to

almost any part of the body

including liver, spleen and bone

marrow.

American Cancer Society estimated new

cancer cases in men and women in 2016.

INDIAN TRADITIONAL MEDICINE

India is known for

its traditional

medicinal systems

Ayurveda, Siddha,

and Unani.

Plant selected

Garcinia morella

fruit Dysentery

Gastritis Inflammatory

diseases

Dried samples

stored for years

Antioxidant

Plant Samples

washed cut and

dried

Dried samples

grounded to

fine powder

Dried powder

Methanol

Evaporate the

extracts by rotar

evaporator

Store the dried

extracts in -20ºC Extraction by

maceration

Preparation of Extracts

IN VITRO ANTIOXIDANT ACTIVITY OF GF, GB AND GL EXTRACTS

A) DPPH radical scavenging activity

B) Reducing power activity

C) Lipid peroxidation inhibition

activity of methanol extracts of G.

morella fruit (GF), bark (GB), leaf

(GL) and standard BHT.

DPPH radical scavenging activity

Reducing power activity

Lipid peroxidation inhibition

In vitro cytotoxic effect of G morella fruit(GF), bark(GB) and leaf (GL)

extracts on Dalton’s ascites lymphoma (DLA)

GF50

GF10

0

GF25

0

GF50

0

GL50

GL10

0

GL25

0

GL50

0

GB50

GB10

0

GB25

0

GB50

0

0

20

40

60

80

100

%In

hib

itio

n

In vitro cell viability of DLA cells

monitored by A) MTT assay and C)

FACS analysis after 3 h treatment of

DLA cells with different concentrations

of GF, GB and GL and B) trypan blue

dye exclusion assay of DLA cells on

treatment with GF at different

concentrations for 3h. All the results are

expressed in mean ± SD (n=3).

A B

C

MTT assay

FACS analysis

Trypan blue

IN VIVO EXPERIMENTAL DESIGN

Group I : Normal animals without inoculation with DLA cells.

Group II : DLA animals + Vehicle control (Fed with 0.3% CMC)

Group III : DLA animals + 10 mg/ kg Cyclophosphamide

Group IV : DLA animals + 100 mg/kg GF methanolic extract

Group V : DLA animals + 200 mg /kg GF methanolic extract

Group VI : DLA animals + 200 mg/kg GL methanolic extract.

Group VII: DLA animals + 200 mg/Kg GB methanolic extract

Animals were injected with

DLA cells (0.2 ml of 2×106

cells/mouse)

After 24

hours

On 10th day of

Drug

administration

animals were

sacrificed and

blood , serum

and liver tissues

collected

Drug

administration

by oral gavage

Haematological Parameters Biochemical Parameters

GF (200mg/kg) restored Haematological and

Biochemical parameters of DLA induced mice

EFFECT OF EXTRACTS ON LIFESPAN OF DALTON’S ASCITES

LYMPHOMA INDUCED MICE

Effect of the G. morella extracts on longevity

of Daltons lymphoma induced mice is

represented by Kaplan meir curve

GF (200mg/kg)

significantly

increases (66 %)

lifespan of DLA

induced animals

DLA DLA+GF 200

A B C

Effect of GF treatment on neovascularisation

The figure (A) , (B) and (C) represents the inner peritoneum lining of

untreated, Standard treated and GF 200 mg/kg treated DLA induced

animals respectively.

D E F

Effect of GF treatment on liver histology

The figures (D), (E) and (F) is the liver histology of DLA induced

untreated , Standard treated and GF200 mg/kg treated animals.

GF EXTRACT INDUCES APOPTOSIS OF DLA CELLS

gf 1

50

gf 2

50

0

1

2

3

g f 1 5 0

g f 2 5 0

fo

ld

c

ha

ng

e in

c

as

pa

se

3

Fold change in level of Caspase3

enzyme of DLA cells upon

treatment with GF150 and GF250

µg/ml

DNA Fragmentation assay. Lane 1) 200bp DNA ladder

Lane 2) DLA +GF50,

Lane 3) DLA + GF 100,

Lane 4) DLA +GF150,

Lane 5) DLA +GF200,

Lane 6) DLA untreated cells

Caspase 3 level DNA fragmentation

Morphology of DLA cells on treatment with GF for 3 hours stained with

AO/EtBr A) Untreated DLA cells B) DLA cells treated with GF150 µg/ml C)

DLA cells treated with GF250 µg/ml

SEM images of DLA cells on treatment with GF for 3 hours A)Untreated DLA

cells B)DLA cells treated with GF150 µg/ml C)DLA cells treated with GF250

µg/ml

Morphological changes of DLA cells on treatment with GF

A B C

CONCLUSION

Garcinia morella fruit methanolic extract has highest in vitro

antioxidant activity in comparison to leaf and bark extracts.

G. morella fruit methanolic extract possess significant in vitro and

in vivo anticancer activity against T cell lymphoma.

G. morella fruit methanolic extract shows apoptotic effect on

DLA cells.

Treatment of DLA induced mice with GF(200mg/kg) also

significantly inhibited neovascularisation in the peritoneum of the

mice and restored the liver histology.

Thank you

bhaswatichoudhury@ymail.com